BPS2026 – Cell-free synthesis and SMALP solubilization for functional analysis of membrane proteins

Membrane proteins play critical roles in biology and medicine but remain challenging to express and stabilize for biochemical and biophysical study. Here, we demonstrate an integrated approach that couples cell-free protein synthesis (CFPS) with styrene-maleic acid lipid particles (SMALPs) to enable direct solubilization and functional reconstitution of membrane proteins without detergents. Using CFPS, we rapidly produced full-length receptors and channels and applied SMALPs to encapsulate and preserve the activity of a membrane-bound protease and a porin within native-like bilayer environments. This workflow supports structural and mechanistic analyses, including electrophysiology, spectroscopy, and binding assays, while maintaining protein folding and activity. By bridging de novo synthesis with detergent-free solubilization, the CFPS-SMALP platform expands experimental access to otherwise intractable systems and establishes a versatile foundation for biochemical and biophysical investigations of diverse membrane proteins.